CN1974554A - Cyclin imide peptidyl metalloprotease inhibitor and its application - Google Patents
Cyclin imide peptidyl metalloprotease inhibitor and its application Download PDFInfo
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Abstract
The present invention provides one kind of powerful peptide-like metalloprotease inhibitor, which exhibits obvious selectivity between endopeptidase and exopeptidase and may be used in treating diseases with abnormal active expression of metalloprotease effectively. Specifically, the present invention relates to the peptide-like compound in the general expression as shown, and its optical isomers, pharmaceutically acceptable salts, solvates and medicine precursor. The present invention also relates to the medicine composition containing the peptide-like compound and its medicinal use.
Description
Technical field
The present invention relates to selectivity that a class has the cyclin imide peptidyl skeleton suppress the metalloprotease effect the class peptide compounds preparation method, activity test and contain the composition of such peptide compounds, and the purposes of these compositions.
Background technology
1, matrix metalloproteinase (MMPs)
MMPs is the endopeptidase that a class relies on calcium ion and zine ion, and the activity of MMPs is by the strict control of the secretion level of gene expression dose and activation of zymogen/supressor.MMPs plays an important role in extracellular matrix degradation, tissue reconstruction process.In a lot of pathologic processes, in the growth and transfer as sacroiliitis, tissue fester, malignant tumour, MMPs has also played vital role.
28 members (Szabo, K.A.et al.Clinical andApplied Immunology Reviews.2004,4 of MMPs family in Mammals, have been found at present, 295), according to its structure, specific substrate and different cell positions are divided into different hypotypes, comprise kind of a collagenase (MMP-1,-8 ,-13 ,-18), 2 kinds of gelatinase (MMP-2,-9), 3 kinds of extracellular matrix degrading enzymes (MMP-3 ,-10,-11), 6 kinds of membranous type-matrix metalloproteinases (MMP-14 ,-15 ,-16,-17,-24 ,-25), and other are unclassified as stromlysin (MMP-7 and-26) and scavenger cell metallic elastic albumen (MMP-12) etc.Wherein gelatinase (MMP-2 and-9) has been proved to be closely related in the poor prognosis of the malignant phenotype of invasive tumor and cancer patient, and they have participated in the invasion and attack of tumour cell to basilar membrane, matrix, to penetrating of vessel wall, and the transfer of tumour cell.Recent study shows, MMPs also with the growth and the associated angiogenesis of primary tumor and secondary tumor, even the tumour birth process also played a driving role.Therefore, aiming is that the therapeutic strategy of action target spot also develops rapidly with these enzymes, and the MMPs inhibitor has become the focus in the cancer treatment drugs research.
The example of available MMPs inhibitor for treating comprises: rheumatoid arthritis (Mullins, D.E.; Et al.Biochim.Biophys.Acta.1983,695,117); Osteoarthritis (Henderson, D.; Et al.Drugs ofthe Future, 1990,15,495); Cancer; Tumour cell shifts (Deryugina, E.I.; Et al.Cancer Metastasis Rev.2006,25,9); Multiple sclerosis (Rosenberg, G.A.et al.Ann Neurol.2001,50,431); And various tissue ulcers or tissue ulcer's venereal disease disease.As the ulcer that occurs in cornea may be because of due to the alkali burn, or because of infecting due to pseudomonas aeruginosa, Acanthamoeba, herpes simplex and the vaccinia virus.
With metal proteinase activity excessively is that other examples of the illness of feature comprise periodontopathy, epidermolysis bullosa, heating, inflammation and scleritis (Cf.Decicco, et al WO95/29892).
2, Aminopeptidase N
Aminopeptidase N (APN/CD13) participates in the degraded of substrate N terminal amino acid for depending on the metal exopeptidase of zine ion.Under the normal condition, APN is extensive low expression level in tissues such as mammiferous kidney, small intestine and central nervous system, participates in the physiological regulation of body.Studies have shown that Aminopeptidase N plays an important role in tumour generation, immunologic function adjusting and virus infection.1) APN is at the tumor cell surface high level expression.This enzyme can make the main component degraded of extracellular matrix (ECM), has destroyed the natural cover for defense of body, promotes invasion by tumor cells, growth and transfer, participates in the tumour neovascularity and generates.(Saiki，I.；et al.Int.J.Cancer.，1993，54，137；Sato，Y.Biol.Pharm.Bull.，2004，772，776)。2) APN has also participated in the inflammatory reaction that the T lymphocyte relies on simultaneously in granulocyte and lymphocytic cell surface great expression; Can also be expressed in the antigen presenting cell surface, degraded immunologic active material (as interleukin-8); The T cell that major histocompatibility complex II type (MHC-II) the Adhesion Antigen determinant of processing of participation antigen and cell surface relies on is to antigenic identification, reduced the recognition capability of T cell, weakened scavenger cell and NK cell identification and kill capability simultaneously, immunity of organisms is descended tumour cell.3) APN plays an important role in upper respiratory tract infection (as: SARS) and acute enteritis as the acceptor on human corona virus HCoV-229E and Transmissible gastroenteritis virus (TGEV) surface, and its relevant (Delmas of activity with enzyme of playing a role, B., et al.Nature, 1992,357,417; Yeager, C.L.; Etal.Nature, 1992,357,420).4) APN participates in the process that the HIV virion enters host cell.Studies show that the APN activity is higher than the healthy volunteer far away in patient's body of infected by HIV.When HIV-1 invasion host cell, the APN of high expression level can make the chemokine fMLP of HIV-1 auxiliary receptor CCR 5 desensitization by degraded, thereby reduces the natural immunity function of cell, and makes the CCR5 enhanced sensitivity, promotes that virus enters host cell.(Shen W, Li B, et al.Blood, 2000,96 (8), 2887; Shipp MA, et al.Blood, 1993,82 (4), 1052) 5) APN participates in the degraded of endogenous analgesic matter endorphin and enkephalin, thus cause the excessive release of P material, cause pain.6) APN degraded Angiotensin, adjusting (Mitsui, the T. of participation body blood pressure; Et al.Biol. Pharm.Bull., 2004,27,768.).
Since the nineties, develop many MMPs inhibitor, great majority are the analogue of peptide or peptide, degraded to enzyme is relatively more responsive, in addition because great majority have backbone, therefore contained rotatable singly-bound number is many, and to bad with the member's selectivity in the family, this also is most of MMPs inhibitor at the reason place that clinical stage is had one shot.Inhibitor at APN mostly is natural product in addition, the medicine ubenimex of a unique listing has the class dipeptides structure that contains beta-amino acids, be used for leukemic treatment as immunostimulant at present, but owing to be to separate to obtain from the nutrient solution of the netted streptomycete of olive (Streptomycesolivorecticuli), it is limited to originate.The inhibitors of metalloproteinase of being reported in the document is not considered the selective problems to the two (endopeptidase and exopeptidase) at present, does not therefore do deep discussion from the mechanism of action aspect.
As everyone knows, because it is little that peptide molecule generally all exists solubleness, bioavailability is low, metabolic stability is poor, very easily limited its clinical application by shortcoming such as various peptide enzyme liberating that extensively exist in the body, class peptide (Peptidomimetics, have another name called peptide analogs or peptide mimics) become novel antitumor owing to himself advantage is expected to replace polypeptide, the strategy of antiviral design and development: on the one hand, the class peptide has the intrinsic activity of substrate, can improve selectivity and usefulness simultaneously by the active centre activity of coming inhibitory enzyme of identification enzyme to target site; In addition, it is difficult by the peptide enzyme liberating that class peptide and native peptides class substrate exist structural difference, biologically stable and availability height, and the long action time of compound; On the other hand, the class peptide can be simulated the native conformation of part, is complementary with the combining site of acceptor, brings into play the effect of its agonist or antagonist.In a word, class peptide layout strategy provides a valid approach for the new drug design and development, also will become antitumor, inverase hot of research and development field simultaneously.
The present invention combines MMP and APN and studies, because matrix metalloproteinase and Aminopeptidase N all belong to zine ion dependency lytic enzyme, and all participate in the degraded of extracellular matrix main component IV Collagen Type VI enzyme and the generation of tumour neovascularity, difference only is that the former is the secretor type endopeptidase, and the latter is the membranous type exopeptidase.When the design compound, introduced class peptide and conformation restriction strategy among the present invention, can improve identification and selectivity on the one hand, improved stability on the other hand lytic enzyme to target.Relate to the selective problems of designed class peptide compounds in this patent, and be expected to develop respectively the specificity selective depressant by optimization to class peptide compounds structure to both.Designed class peptide compounds is in micromole's rank and embodies special selectivity (IC the inhibition activity of MMP among the present invention
50APN/IC
50MMP is near 150), be directed to the APN screening active ingredients in addition and find that its activity of several drug molecules is equivalent to the ubenimex of present unique listing.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, a kind of cyclin imide peptidyl metalloprotease inhibitor and application thereof are provided, the preparation method of cyclin imide peptidyl metalloprotease inhibitor also is provided.
The present invention is a starting raw material with optical purity L-glutamic acid or aspartic acid, we are stitched to gallic acid, coffic acid, toluylic acid, forulic acid and NSAID (non-steroidal anti-inflammatory drug) organic acid (indole-3-acetic acid, diclofenac) on L-glutamic acid or the aspartic acid parent nucleus on this basis, form dicarboxylic acid and then process cyclization, be converted into the cyclin imide skeleton of conformation restriction, improve selectivity target.And these organic acids self have anti-tumor activity, can strengthen to press down enzymic activity.Therefore amino acid can improve the consistency of compound to tissue owing to have natural character.In addition; can be raw material also with optically pure glutamine or l-asparagine; by the synthetic key intermediate of steps such as Boc or Cbz protection, cyclization, replacement; deprotection then is again by obtaining different series class dipeptides or class tripeptides with the different acyl chlorides (as gallic acid, coffic acid, toluylic acid, forulic acid, NSAID (non-steroidal anti-inflammatory drug) organic acid) and the amino acid derivative of biologically active.Purpose all is for avidity that strengthens compound and enzyme or acceptor and metabolic stability.
The present invention designs and has synthesized the inhibitors of metalloproteinase that a class has the brand new parent nucleus.But in vitro tests shows its acellular cytotoxic activity but embodies remarkable vitro enzymic activity, is expected to become the anticancer drug candidate of a class non-cytotoxicity class.
Technical scheme of the present invention is as follows:
Class peptide compounds with general formula I, with and optical isomer, diastereomer and racemic mixture, its pharmacy acceptable salt, solvate or prodrug.
Wherein,
R
1Be aryl, heteroaryl, aryl C1-6 alkyl, heteroaryl C1-9 alkyl; aryl C2-6 thiazolinyl, heteroaryl C2-6 thiazolinyl, aryl C2-6 alkynyl; heteroaryl C2-6 alkynyl, the C1-6 alkyl, the N end is protected or is not protected natural or the alpha-non-natural amino acid derivative; optional by one or more following groups replacements: hydroxyl, halogen, nitro; cyano group, halogen C1-8 alkyl, C1-8 alkoxyl group; the C1-6 alkyl-carbonyl, C1-8 carbalkoxy, aryl C1-8 carbalkoxy.
R
2Be hydrogen, aryl, heteroaryl, aryl C1-6 alkyl, heteroaryl C1-9 alkyl, aryl C2-6 thiazolinyl, heteroaryl C2-6 thiazolinyl, aryl C2-6 alkynyl, heteroaryl C2-6 alkynyl, and various natural or alpha-non-natural amino acid side chains, optional by one or more following groups replacements: halogen, nitro, cyano group, halogen C1-8 alkyl, C1-8 alkoxyl group, the C1-6 alkyl-carbonyl, the C1-8 carbalkoxy.
R
3Be OH, NHOR
10(R here
10Be hydrogen or C1-8 alkyl), C1-8 alkoxyl group, NHR
11(R here
11Be the C1-8 alkyl, aryl, heteroaryl, aryl C1-6 alkyl, heteroaryl C1-9 alkyl, aryl C2-6 thiazolinyl, heteroaryl C2-6 thiazolinyl, aryl C2-6 alkynyl, heteroaryl C2-6 alkynyl, NHCH (R
2) COR
4(R here
4Be OH or NHOR
10), optional by one or more following groups replacements: halogen, nitro, cyano group, halogen C1-8 alkyl, the C1-8 alkoxyl group, the C1-6 alkyl-carbonyl, C1-8 alcoxyl carbonyl,
N is 1 or 2,
* be that steric configuration is S or R optical purity or its raceme.
Preferably, R
1Be aryl, aryl C1-6 alkyl, aryl C2-6 thiazolinyl, the non-steroidal anti-inflammatory drugs organic acid, the N end is protected or do not protected natural or the alpha-non-natural amino acid derivative, and is optional by one or more following groups replacements: hydroxyl, halogen, nitro, cyano group, C1-8 alkoxyl group;
R
2Be hydrogen, and various natural or alpha-non-natural amino acid side chain;
R
3Be OH, OCH
3, NHCH (R
2) COR
4, R here
4Be OCH
3Or NHOH.
Above-mentioned class peptide compounds (I) specifically comprises following compound:
(S)-methyl-2-(2-(2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) acetic ester
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3 Methylbutanoic acid ester
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-4-methylpent acid esters
(2S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3 methylvaleric acid ester
(S)-methyl 1-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) ethanoyl) tetramethyleneimine-2-carboxylicesters
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) propionic ester
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-5-(3-nitro guanidine radicals) valerate
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-4-(methylthio group) butyric ester
(S)-methyl 6-(benzyloxy carbonyl amide group)-2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) capronate
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-(4-hydroxybenzyl) propionic ester
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-(1H-indol-3-yl) propionic ester
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-(1H-imidazoles-5-yl) propionic ester
(R)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-phenylpropionic acid ester
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-hydroxy propionate
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-phenylpropionic acid ester
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-mercaptopropionic acid ester
(S)-dimethyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) phthalic ester
(S)-methyl 3-chloro-2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) propionic ester
(S)-methyl 3-(2-(2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) propionic ester
(S)-methyl 2-(2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetic ester
(S)-and N-(1-(2-(azanol base)-2-oxygen ethyl)-2,6-dioxopiperidin-3-yl)-3,4,5-trimethoxy aniline.
The class peptide compounds intermediate of above-mentioned general formula (I) is: 3,4, and the 5-trimethoxybenzoic acid; 3,4, the 5-trimethoxy-benzoyl chloride; (S)-2-(3,4,5-trimethoxy anilino) pentanedioic acid; (S)-and N-(2,6-dioxy-tetrahydrochysene-2H-pyrans-3-yl)-3,4, the 5-trimethoxy-benzamide; (S)-2-(2,6-dioxy-3-(3,4,5-trimethoxy benzamido) piperidines-1-yl) acetate, (S)-tertiary butyl-2,6-dioxopiperidin-3-aminocarbamic acid ester, (S)-ethyl-2-(3-(t-butoxycarbonyl amino)-2,6-dioxopiperidin-1-yl) acetic ester, (S)-benzyl-2,6-dioxopiperidin-3-aminocarbamic acid ester, (S)-ethyl 2-(3-(benzyloxycarbonyl amino)-2,6-dioxopiperidin-1-yl) acetic ester.
The preparation method of above-mentioned class peptide compounds is a raw material with the optical purity acidic amino acid, condensation under alkaline condition, and the compound of gained passes through dehydration condensation again, nucleophilic substitution, polypeptide condensation reagent condensations etc. are reacted final separation and purification and are got; Or be raw material with optically pure glutamine or l-asparagine; by the synthetic key intermediate of steps such as Boc or Cbz protection, cyclization, replacement; deprotection then, class dipeptides or class tripeptides again by obtaining different series with the different acyl chlorides and the amino acid derivative of biologically active.
These class peptide compounds comprise matrix metalloproteinase or Aminopeptidase N at prevention or treatment and metalloprotease, the application of the medicine of the mammalian diseases that active unconventionality expression is relevant.
Described and the related mammalian disease metal proteinase activity unconventionality expression comprises: inflammation, cancer, multiple sclerosis, various tissue ulcers or tissue ulcer's venereal disease disease, periodontopathy, epidermolysis bullosa, leukemia etc.Therefore, the invention still further relates to the pharmaceutical composition that contains (I) structural compounds.
A kind of pharmaceutical composition comprises (1) above-mentioned each class peptide compounds and (2) one or more pharmaceutically acceptable carriers or vehicle.
In addition, the present invention also comprises a kind of mammiferous pharmaceutical composition of orally give that is suitable for, and comprises (1) above-mentioned arbitrary class peptide compounds and (2) pharmaceutically acceptable carrier, optional (3) one or more the pharmaceutically acceptable vehicle that comprise.
In addition, the present invention comprises that also a kind of parenteral that is suitable for gives mammiferous pharmaceutical composition, comprises (1) above-mentioned arbitrary class peptide compounds and (2) pharmaceutically acceptable carrier, optional (3) one or more the pharmaceutically acceptable vehicle that comprise.
Class peptide and conformation layout strategy have been adopted in the design that the present invention contains the compound of general formula (I) first.Class peptide and conformation restriction strategy have been widely used in design and development field antiviral, antitumor drug, its structure is made of the structure that is similar to peptide natural or alpha-non-natural amino acid, but overall conformation is by being different from natural peptide material, on the one hand, the class peptide has the intrinsic activity of substrate, can improve selectivity and usefulness simultaneously by the active centre activity of coming inhibitory enzyme of identification enzyme to target site; In addition, it is difficult by the peptide enzyme liberating that class peptide and natural peptide matters exist structural difference, biologically stable and availability height, and the long action time of compound.
Detailed Description Of The Invention
Used definition and term
Term and definition implication used herein is as follows:
" assorted alkyl " refers to saturated or unsaturated, carbon atoms and at least one heteroatomic chain, and wherein any one heteroatoms is non-conterminous.Contain 2-15 atom (carbon atom) in the assorted alkyl, preferably contain 2-10 atom.Assorted alkyl can be direct-connected or side chain, replacement or unsubstituted.
" aryl " is meant the aromatic carbocyclic group.Preferred aromatic ring contains 6-10 carbon atom.
" halogen ", or " halogen " comprise fluorine, chlorine, bromine or iodine, preferably not fluorine and chlorine.
" cycloalkyl " is replacement or unsubstituted, saturated or undersaturated cyclic group, and it contains carbon atom and/or one or more heteroatoms.This ring can be monocycle or condensed ring, the ring system of bridged ring or volution.Monocycle has 3-9 atom usually, preferably has 4-7 atom, and many rings contain 7-17 atom, preferably contain 7-13 atom.
" heteroaryl " is aromatic heterocycle, can be monocycle or bicyclic radicals.Preferable heteroaryl comprises, thienyl for example, and furyl, pyrryl, pyridyl, pyrazinyl, thiazolyl, pyrimidyl, quinolyl and tetrazole base, benzothiazolyl, benzo furan are fed base, indyl etc.
" pharmacy acceptable salt " is meant that formula (I) compound has curative effect and nontoxic salt form.It can form anion salt by arbitrary acidic-group (as carboxyl), or forms cationic salts by arbitrary basic group (as amino).A lot of such salt known in the art.Go up the cationic salts that forms at any acidic-group (as carboxyl), or go up the anion salt that forms at any basic group (as amino).These salt are known in the art by many formulas, comprise the salt and the organic salt (as ammonium salt) of basic metal (as sodium and potassium) and alkaline-earth metal (as magnesium and calcium) as cationic salts.Also can obtain anion salt easily by (I) that uses corresponding acid treatment alkaline form, such acid comprises mineral acid such as sulfuric acid, nitric acid, phosphoric acid etc.; Or organic acid such as acetate, propionic acid, oxyacetic acid, 2 hydroxy propanoic acid, 2-oxo propionic acid, oxalic acid, propanedioic acid, succsinic acid, toxilic acid, fumaric acid, oxysuccinic acid, tartrate, 2-hydroxyl-1,2,3-the third three acid, methylsulfonic acid, ethyl sulfonic acid, benzene methanesulfonic acid, 4-toluene sulfonic acide, cyclohexyl-sulfinic acid, 2 hydroxybenzoic acid, 4-amino-2-hydroxybenzoic acid etc.These salt formulas those of skill in the art know, and those skilled in the art can prepare any salt that this area knowledge is provided.In addition, those of skill in the art can get certain salt according to solubleness, stability, easy preparation etc. and give up another kind of salt.The mensuration of these salt and optimization are in those of skill in the art's experience scope.
" solvate " is the title complex that solute (as inhibitors of metalloproteinase) and solvent (as water) are combined to form.Referring to J.Honig etc., The Van Nostrand Chemist ' s Dictionary, p.650 (1953).The pharmaceutically acceptable solvent that the present invention adopts comprises bioactive those solvents of not disturbing inhibitors of metalloproteinase (solvent known to for example water, ethanol, acetate, the N, dinethylformamide, dimethyl sulfoxide (DMSO) and this those skilled in the art or that determine easily).
" optical isomer " used herein, " enantiomorph ", " diastereomer ", " raceme " etc. have defined the form of The compounds of this invention or all possible steric isomer of its physiological derivative.Unless indication is arranged in addition, the chemical name of The compounds of this invention comprises the mixture of all possible stereochemical form, affiliated mixture comprises all diastereomers and the enantiomorph of basic structure molecule, and the single isomeric forms of the The compounds of this invention of substantially pure, promptly wherein contain and be lower than 10%, preferably be lower than 5%, particularly be lower than 2%, most preferably be lower than other isomer of 1%.The various stereoisomer forms of class peptide compounds of the present invention all obviously are contained in the scope of the present invention.
The form of all right other protected form of formula (I) class peptide compounds or derivative exists, and these forms will be apparent to those skilled in the art, and all should be contained in the scope of the present invention.
Aforesaid substituting group self also can be replaced by one or more substituting groups.Such substituting group is included in C.Hansch and A.Leo, those substituting groups of listing among the Substituent Constants for Correlation Analysis in Chemistry and Biology (1979).Preferred substituted comprises, alkyl for example, thiazolinyl, alkoxyl group, hydroxyl, the oxygen base, nitro, amino, aminoalkyl group (as aminomethyl etc.), cyano group, halogen, carboxyl, carbonylic alkoxy (as carbonyl oxyethyl group etc.), sulfenyl, aryl, cycloalkyl, heteroaryl, Heterocyclylalkyl (as piperidyl, morpholinyl, pyrryl etc.), imino-, hydroxyalkyl, aryloxy, arylalkyl, and combination.
Synthetic
Target compound is synthetic through following route.
In brief, the preparation method of the class peptide compounds of above-mentioned general formula (I), the present invention is a starting raw material with optical purity L-glutamic acid or aspartic acid, we are stitched to gallic acid, coffic acid, toluylic acid, forulic acid and NSAID (non-steroidal anti-inflammatory drug) organic acid (indole-3-acetic acid, diclofenac) on L-glutamic acid or the aspartic acid parent nucleus on this basis, form dicarboxylic acid and then process cyclization, be converted into the cyclin imide skeleton of conformation restriction, improve selectivity target.And these organic acids self have anti-tumor activity, can strengthen to press down enzymic activity.Therefore amino acid can improve the consistency of compound to tissue owing to have natural character.In addition; can be raw material also with optically pure glutamine or l-asparagine; by the synthetic key intermediate of steps such as Boc or Cbz protection, cyclization, replacement; deprotection then is again by obtaining different series class dipeptides or class tripeptides with the different acyl chlorides (as gallic acid, coffic acid, toluylic acid, forulic acid, NSAID (non-steroidal anti-inflammatory drug) organic acid) and the amino acid derivative of biologically active.Purpose all is for avidity that strengthens compound and enzyme or acceptor and metabolic stability.Concrete synthetic route designs in conjunction with the state of the art according to certain concrete compound that will prepare.Be example with gallic acid cyclin imide peptidyl derivative below, the synthetic route of this compounds of declaratives and preparation method.
Reagent: a.1) methyl-sulfate, sodium hydroxide 2) hydrochloric acid; B. sulfur oxychloride, benzene; C.L-L-glutamic acid, yellow soda ash, 2) hydrochloric acid; D. diacetyl oxide, 55-60 ℃; E. glycine, N, dinethylformamide, 110 ℃, 5h; F. amino acid methyl ester hydrochloride, 1-ethyl-3-(3-dimethylamine propyl) carbodiimide hydrochloride (EDCI), I-hydroxybenzotriazole, methylene dichloride/dimethyl sulfoxide (DMSO); G.1) azanol, potassium hydroxide, methyl alcohol 2) boron tribromide, methylene dichloride ,-78 ℃; H. sulfur oxychloride, methyl alcohol; I. azanol, potassium hydroxide, methyl alcohol.
Use Me
2The free hydroxyl group of SO4 protection gallic acid; then with the condensation under alkaline condition of L-L-glutamic acid; (the S)-2-(3 of gained; 4; 5-trimethoxy anilino) pentanedioic acid, dehydration condensation in diacetyl oxide, the product of gained in non-protonic solvent with the glycine nucleophilic substitution; again in DCM or THF through polypeptide condensing agent EDCI condensation, separation and purification promptly gets final product.
Those skilled in the art can change to improve yield above-mentioned steps; they can determine the synthetic route according to the ABC of this area; as the selective reaction thing, solvent and temperature, thus can improve yield with the generation of avoiding side reaction by using various GPF (General Protection False bases.These conventional guard methods can be referring to for example T.Greene, Protecting Groups inOrganic Synthesis.
Obviously, above-mentioned route is that stereoselectivity is synthetic, can also can prepare its optically active class peptide compounds by above-mentioned route.For example raw material L-L-glutamic acid is replaced with its optical isomer (R configuration).Those skilled in the art can obtain various other isomer of cyclic imide derivative easily, and can be by conventional separation means purifying, as chirality salt or chirality chromatography column etc.
MMPs suppresses active test description in Vijaykumar, M.B. etc., and Matrix Biol.2000 is in 19,26.Succinyl gelatin has proved can be by gelatinase (comprising MMP-2 ,-9) hydrolysis, and the height of the free amine group concentration that peptide bond hydrolysis produces is proportionate with the enzymic activity size.Free amine group in the succinyl oxide protection gelatin, the uncle's ammonia and 2,4 that exposes after the hydrolysis, 6-trinitro-benzene-sulfonic acid (TNBS) reaction solution is determined amino content by the optical density that detects the 450nm place, thereby determines the activity of gelatinase.
APN suppresses active test description in Lejczak, and .Biochemistry such as B are in 1989,28,3549.Substrate L-leucyl-p-N-methyl-p-nitroaniline is degraded by APN, be created in the p-N-methyl-p-nitroaniline that 405nm has absorption, and the size of the concentration of p-N-methyl-p-nitroaniline and enzymic activity is proportionate.Determine the content of p-N-methyl-p-nitroaniline by the optical density that detects the 405nm place, thereby determine the activity of aminopeptidase, reflect that indirectly inhibitor suppresses the size of degree to enzymic activity.
The class peptide compounds of general formula (I) external presses down enzyme test proves that such peptide compounds is a kind of cyclin imide peptidyl metalloprotease inhibitor
Therefore cyclic imide derivative of the present invention spatially is complementary with the MMP activities site, has shown higher inhibition activity external.And it can be metabolized to active fragments in vivo, as gallic acid-derivate, still has anti-tumor activity, has therefore also shown higher anti-tumor activity in vivo.
Preparation, pharmaceutical composition, dosage and taking
Cyclic imide derivative of the present invention can free form or is existed with salt form.Pharmacy acceptable salt of the known chemical compound lot type of one skilled in the art and preparation method thereof.Pharmacy acceptable salt comprises conventional avirulent salt, comprises such compound alkali and quaternary ammonium salt inorganic or that organic acid forms.
Compound of the present invention can form hydrate or solvate.The one skilled in the art known with compound formed hydrate or form the method for solvate when in solution, concentrating during with the water freeze-drying with appropriate organic solvent.
The present invention comprises the medicine that contains the therapeutic dose The compounds of this invention and the pharmaceutical composition of one or more pharmaceutically acceptable carriers and/or vehicle.Carrier comprises as salt solution, buffer saline, and glucose, water, glycerine, ethanol and their binding substances are hereinafter discussed in more detail.If desired, said composition can also comprise wetting agent or emulsifying agent in a small amount, or the pH buffer reagent.Said composition can be liquid, suspension, emulsion, tablet, pill, capsule, extended release preparation or powder.Said composition can be mixed with suppository with traditional tamanori and carrier such as triglyceride.Oral preparations can comprise the mannitol of standard vector such as medicine grade, lactose, starch, Magnesium Stearate, soluble saccharin, Mierocrystalline cellulose and magnesiumcarbonate or the like.Preparation and deciding optionally, preparation can design mixing, granulation and compression or solvent components.In another approach, said composition can be mixed with nano particle.
The pharmaceutical carrier that uses can for, for example, solid or liquid.
The typical solid carrier comprises lactose, terra alba, sucrose, talcum, gel, agar, pectin, gum arabic, Magnesium Stearate, stearic acid or the like.Solid carrier can comprise that one or more may be simultaneously as sweetener, lubricant, solubilizing agent, suspension agent, filler, glidant, compression aid, the material of tackiness agent or tablet-disintegrating agent; It can also be an encapsulating material.In powder, carrier is pulverizing solid, and it mixes with pulverizing activeconstituents.In tablet, Mars composition and the carrier with necessary compression property are with suitable mixed, with the shape and the size compression of needs.Powder and tablet preferably comprise 99% activeconstituents at the most.Suitable solid carrier comprises, for example, and calcium phosphate, Magnesium Stearate, talcum, sugar, lactose, dextrin, starch, gel, Mierocrystalline cellulose, methylcellulose gum, sodium carboxymethyl-cellulose, polyvinylpyrrolidone alkane ketone, low melt wax and ion exchange resin.
Exemplary of liquid carriers comprises syrup, peanut oil, and sweet oil, water, or the like.Liquid vehicle is used to prepare solution, suspension, emulsion, syrup, the composition of tincture and sealing.Activeconstituents can dissolve or be suspended in pharmaceutically acceptable liquid vehicle such as water, organic solvent, the mixture of the two or pharmaceutically acceptable oils or fat.Liquid vehicle can comprise other suitable medicated premix such as solubilizing agent, emulsifying agent, and buffer reagent, sanitas, sweetener, sweetener, suspension agent, thickening material, pigment, viscosity modifier is stablized shape or osmotic pressure-conditioning agent.The suitable example that is used for the liquid vehicle of oral and administered parenterally comprises that water (partly comprises as above-mentioned additive, derivatived cellulose for example, the preferably carboxymethyl cellulose sodium salt solution), alcohol (comprises monohydroxy-alcohol and polyvalent alcohol, and oils (for example fractionated coconut oil and peanut oil) ethylene glycol for example) and their derivative.The carrier that is used for administered parenterally can also be grease such as ethyl oleate and sec.-propyl myristate.Aseptic liquid vehicle is used for the aseptic fluid composition of administered parenterally.The liquid vehicle that is used for pressurized compositions can be halohydrocarbon or other pharmaceutically acceptable propelling agents.Sterile solution or aaerosol solution composition of liquid medicine can be used for, for example, and intravenously, intramuscular, intraperitoneal or subcutaneous injection.But single pushes or injection gradually during injection, goes into 30 minutes the interior perfusion of passages through which vital energy circulates.This compound can also be with the form oral administration of liquid or solids composition.
Carrier or vehicle can comprise time lag material known in the art, as glyceryl monostearate or distearin, also can comprise wax, ethyl cellulose, Vltra tears, methyl methacrylate or the like.When preparation is used for when oral, generally acknowledge PHOSALPG-50 (phospholipid and 1, the 2-propylene glycol is concentrated, A.Nattermann ﹠amp; Cie.GmbH) 0.01% tween 80 in is used for the preparation of the acceptable oral preparation of other compounds, can be adapted to the preparation of all cpds of the present invention.
Can use medicament forms miscellaneous when giving The compounds of this invention.If the use solid carrier, preparation can be tablet, is placed into powder or piller form or lozenge or lozenge form in the hard capsule.The amount of solid carrier changes to a great extent, but preferably from about 25mg to about 1.0g.If the use liquid vehicle, preparation can be syrup, emulsion, soft capsule, aseptic injectable solution or suspension in the liquid suspension of ampoule or bottle or non-water.
In order to obtain stable water miscible formulation, compound or its pharmacy acceptable salt can be dissolved in the organic or inorganic aqueous acid, 0.3M succsinic acid or citric acid solution.Optionally, the tart derivative can be dissolved in suitable basic solution.If can not get soluble form, compound can be dissolved in suitable cosolvent or their combination.The example of suitable cosolvent like this includes but are not limited to, and concentration range is from the ethanol of 0-60% cumulative volume, propylene glycol, Liquid Macrogol, polysorbate 80, glycerine, polyoxyethylene fatty acid ester, Fatty Alcohol(C12-C14 and C12-C18) or glycerine hydroxy fatty acid ester or the like.
Various release systems are known and can be used for the administration of compound or other various preparations, and these preparations comprise tablet, capsule, and injectable solution, the capsule in the liposome, particulate, microcapsule, or the like.The method of introducing includes, but are not limited to skin, intracutaneous, intramuscular, endoperitoneal, intravenous, subcutaneous, nasal cavity, lung, peridural, eyes and (preferred usually) oral route.Compound can be by administration easily any or that other is suitable, for example by injecting or bolus injection, by epithelium or the mucous membrane circuit (for example, oral mucosa, rectum and intestinal mucosa, or the like) absorb or the support by carrying medicament and can be in other biological promoting agent administration together.Can whole body or topical.Be used for nose, when the treatment of segmental bronchus or lung disease or prevention, preferred route of administration is oral, nasal administration or segmental bronchus smoke substance or atomizer.
Embodiment
The present invention is described further below in conjunction with embodiment, but be not limited thereto.
The preparation of embodiment 1. (S)-2-(3,4,5-trimethoxy anilino) pentanedioic acid (4)
1) 3,4,5-trimethoxybenzoic acid (2).(80g 2mol) is dissolved in the 500ml distilled water NaOH, is cooled to room temperature, adds 3,4, and (0.294mol), the jam-pack stopper is stirred to molten the 5-trihydroxybenzoic acid immediately for gallic acid, 50g.Temperature is lower than 20 ℃ in the control, adds (CH
3)
2SO
4(0.71mol), this moment, temperature can rise to 30～45 ℃, opened the bottle stopper venting frequently for 89g, 67ml.Behind the 20min, add (the CH of second batch of equivalent
3)
2SO
4, this moment, temperature can rise to 40～45 ℃, stirred 10min, heated up and boiled 2h.The NaOH solution that adds 50g 40% again boils 2h, makes the ester saponification that may generate.The cooling reaction solution is used the dilute hydrochloric acid acidifying to room temperature, suction filtration, and the cold water washing filter cake gets crude product 48g.
Making with extra care of crude product.Crude product 4g heating for dissolving in 200ml distilled water, is added the 0.2g gac, boil 20min, filtered while hot is placed the filtrate refrigerator, separates out white, needle-shaped crystals, filters the drying 3.4g that weighs, 168～171 ℃ of mp.
2) 3,4,5-trimethoxy-benzoyl chloride (3).3,4, (21.2g 0.1mol) is dissolved in the 250ml benzene 5-trimethoxybenzoic acid, adds 25ml SOCl
2, reflux 4h.Stopped reaction, the evaporated under reduced pressure solvent adds benzene 50ml dissolving back evaporate to dryness again, repeats twice, to eliminate SOCl
2It is standby to add the dissolving of 200ml benzene.
3) (S)-2-(3,4,5-trimethoxy anilino) pentanedioic acid (4).(19.6g, 0.185mol) (16.9g 0.115mol) is dissolved in the 150ml water anhydrous sodium carbonate, slowly drips the benzole soln cryosel bath simultaneously of above-mentioned acyl chlorides under the mechanical stirring, and about 1h adds with L-L-glutamic acid.Stop behind the mechanical stirring 5h stirring, transfer pH to slightly acidic with sour water, standing demix discards the benzene layer.Continue to transfer pH and use chloroform extraction, monitor to water with thin layer simultaneously and have only a point, discard chloroform layer, water continues to transfer pH to 2, separates out a large amount of white flockss, and the refrigerator placement is spent the night.The filtration drying 28.6g that weighs, productive rate 89.3%, mp120～121 ℃.IR(KBr，cm-1)：3299.7，3255.4，2942.5，1744.7，1699.5，1500.7，1235.5，1127.7；
1H-NMR(DMSO-d6，ppm)：12.41(s，2H)，8.53(d，1H，J＝7.2Hz)，7.22(s，2H)，4.41(m，1H)，3.83(s，6H)，3.71(s，3H)，2.35(m，2H)，2.21(m，1H)，1.96(m，1H)；ESI-MS：m/z 341.8。
The preparation of embodiment 2. (S)-2-(2,6-dioxy-3-(3,4,5-trimethoxy benzamido) piperidines-1-yl) acetate (6)
1) (S)-and N-(2,6-dioxy-tetrahydrochysene-2H-pyrans-3-yl)-3,4,5-trimethoxy-benzamide (5).3,4, (10g 2.9mmol), adds the 80ml diacetyl oxide to 5-trimethoxy-benzene carbamylglutamic, 55～60 ℃ of oil bath insulation 5h.Filtered while hot is removed insolubles, adds an amount of anhydrous diethyl ether, and white solid is separated out in cooling.The filtration 5.2g that weighs, productive rate 55%, mp150～152 ℃.IR(KBr，cm-1)：3310.0，2945.1，1777.0，1640.7，1504.2，1239.6，1129.6；ESI-MS：m/z323.8。
2) (S)-2-(2,6-dioxy-3-(3,4,5-trimethoxy benzamido) piperidines-1-yl) acetate (6).Compound 5 (5.0g) is dissolved in 50mlDMF, and the 1.0g glycine grinds the back and adds among the DMF, and 110 ℃ were heated 5 hours.Mixture cooling causes room temperature, falls as isopyknic pH is that placement is spent the night, and separates out a large amount of white crystals in 2 the frozen water.The filtration washing 3.0g that weighs, productive rate 70%.
1H-NMR(DMSO-d6，ppm)：δ12.34(s，1H)，8.88(d，1H)，7.17(s，2H)，4.53(m，1H)，4.42(s，2H)，3.83(s，6H)，3.71(s，3H)，2.18(dt，2H)，2.11(dt，2H)；ESI-MS m/z[M+1]
+381.3。
The preparation of embodiment 3. (S)-methyl-2-(2-(2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) acetic ester (7a).
Compound 6 (1.90g, 5mmol) and HoBt (0.81g 6mmol) is dissolved in 20ml methylene dichloride and 2ml dimethyl sulfoxide (DMSO), ice bath is cooled to 0 ℃, slowly drips EDCI (1.44g, dichloromethane solution 7.5mmol), dropwised in about 1 hour, removed the ice bath stirring at room 2 hours.Add glycine methyl ester hydrochloride under the condition of ice bath (0.75g 6mmol), regulates pH value 7 with triethylamine, treats that ice cube melts automatically, continues to stir and spends the night in batches.Reaction solution is successively with lN hydrochloric acid, 1% yellow soda ash, and the saturated brine washing, dried over sodium sulfate, column chromatography purification obtains final product, productive rate 79%.mp98.7-100.6℃.
1HNMR(400MHz，DMSO-d6，ppm)：δ9.04(t，1H)，8.88(d，1H)，7.17(s，2H)，4.53(m，1H)，4.37(s，2H)，4.16(s，2H)，3.83(s，6H)，3.71(s，3H)，3.67(s，3H)，2.18(dt，2H)，2.11(dt，2H)。ESI-MS m/z[M+1]
+ 452.5。
Embodiment 4. (S)-methyl-2-(2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetic ester (8)
Compound 6 DLs in the 20ml anhydrous methanol, are slowly dripped SOCl under the condition of ice bath
2, after dropwising, remove ice bath, in 35-40 ℃ of stirring, solution becomes must be clarified in about 4 hours.Rotation is steamed and to be desolventized, and adds the 50ml dissolve with methanol again and steams and remove, and after twice, adds 50ml methyl alcohol repeatedly, steams to remove to cause 1/3 volume, and room temperature is placed and separated out a large amount of white plates crystallizations, productive rate.mp 134-137℃；IR(KBr，cm-1)：3423.5，3216.3，2945.7，1743.1，1675.6，1632.8，1343.3，1127.8；
1H-NMR(DMSO-d6，ppm)：δ8.88(d，1H，)，7.17(s，2H)，4.53(m，1H，)，4.44(s，2H)，3.83(s，6H)，3.71(s，3H)，3.67(s，3H)，2.18(dt，2H，)，2.11(dt，2H)；ESI-MS m/z [M+1]
+395.4。
Embodiment 5. (S)-N-(1-(2-(azanol base)-2-oxygen ethyl)-2,6-dioxopiperidin-3-yl)-3,4,5-trimethoxy aniline (9).789mg (2mmol) compound 8 is dissolved in the 7mL anhydrous methanol, adds 1.5mL NH
2The methanol solution of OK, (press Fieserand Fieser, Vol 1, P 478 preparations), stirring at room adds 1.5g silica gel behind the 24h, rotation is steamed and desolventized, and silicagel column reduces pressure on the gained dry powder silica gel, trichloromethane: methyl alcohol (50: 1～5: 1) wash-out, get faint yellow solid 512mg, meet FeCl
3It is red that solution shows, productive rate 64.7%.Fusing point 117.3-118.8 ℃.δ8.88(d，1H)，8.0(d，1H)，7.17(s，2H)，4.53(m，1H)，4.37(s，2H)，3.83(s，6H)，3.71(s，3H)，2.18(dt，2H)，2.11(dt，2H)，2.0(brs，1H，OH).ESI-MS m/z[M+1]
+ 396.5。
Embodiment 6 target compounds suppress gelatinase activity test (In vitro)
Test principle and detailed step are referring to CN 1528745A pyrrolidinyl metalloprotease inhibitor and preparation method thereof.
Experimental result sees Table one.
Embodiment 7 target compounds suppress the activity test (In vitro) of Aminopeptidase N
1 principle: Aminopeptidase N and its substrate (L-leucyl-p-N-methyl-p-nitroaniline) interact, and be created in right-N-methyl-p-nitroaniline that 405nm has absorption, and the size of the concentration of right-N-methyl-p-nitroaniline and enzymic activity are proportionate.Determine the content of right-N-methyl-p-nitroaniline by the optical density that detects the 405nm place, thereby determine the activity of aminopeptidase, reflect that indirectly inhibitor suppresses the size of degree to enzymic activity.
2 materials and methods:
Aminopeptidase N and substrate L-leucyl-p-N-methyl-p-nitroaniline are all available from Sigma company
The preparation of solution:
Damping fluid, preparation pH7.2, the 50mM phosphate buffered saline buffer, room temperature is placed standby.
Aminopeptidase N is dissolved in the solution that is made into 0.2mg/mL in the damping fluid;
Substrate is dissolved in and is made into 0.5mg/mL solution among the DMSO, and each solution refrigerator is placed standby.
3 amino-peptidase activity analyses:
Add above-mentioned Aminopeptidase N solution 10 μ L in 96 orifice plates, substrate solution is supplied 200 μ L with damping fluid.Room temperature is placed 1h, measures absorption value in 405nm wavelength place.
| Numbering | 1 | 2 | 3 | 4 |
| Aminopeptidase N solution (μ L) | 10 | 10 | 10 | 10 |
| Substrate solution (μ L) damping fluid (μ L) optical density | 5 185 0.247 | 10 180 0.408 | 20 170 0.589 | 40 150 0.934 |
Blank group
| Numbering | 1 | 2 | 3 | 4 |
| Substrate solution (μ L) damping fluid (μ L) optical density | 5 195 0.050 | 10 190 0.051 | 20 180 0.062 | 40 160 0.071 |
The optical density of blank group and the no reagent wells optical density in 96 orifice plates (it is 0.035 that no reagent wells absorbs) very nearly the same illustrate that blank organizes and almost do not have absorption.The contrast optical density as can be seen, fixedly the time, substrate solution preferably selects 15 μ L preferably, is 15 μ L so decide substrate at enzyme solution.
4 press down enzyme detects:
Add above-mentioned Aminopeptidase N solution 10 μ L in 96 orifice plates respectively, substrate 15 μ L, the compound 20 μ L of different gradient concentrations, 50mM phosphate buffered saline buffer (pH7.2) is supplied 200 μ L.100% group does not contain inhibitor.Blank group replaces organized enzyme with the enzyme of inactivation, all supplies 200 μ L with damping fluid.Room temperature is placed 1h, measures absorption value in 405nm wavelength place.Calculate inhibiting rate according to following formula:
Inhibiting rate=(100% optical density-compound optical density)/(100% optical density-blank optical density)
According to compound concentrations and corresponding inhibition ratio, utilize origin7.5 software match beneficial effect curve, calculate the IC of each compound
50
Table one, the external enzyme test result that presses down
| Compds | R | IC 50μM a | IC 50(APN)/ IC 50(MMP-2) | |
| APN | MMP-2 | |||
| 6 7a 7b 7c 7d 7e 7f 7g 7h 7i 7j 7k 7l | OH Gly-OMe Val-OMe Leu-OMe Ile-OMe β-Ala-OMe Ala-OMe Arg(NO 2)-OMe Met-OMe Z-Lys-OMe Tyr-OMe Trp-OMe His-OMe | 49.8±5.5 55.4±3.5 21.4±2.6 46.2±3.2 49.3±2.9 53.7±8.4 18.9±4.4 42.0±7.6 20.7±2.7 38.1±3.8 44.8±5.2 43.1±3.2 5.2±2.1 | 4.0±0.5 4.4±1.1 1.0±0.2 3.8±0.5 4.3±0.4 2.2±0.6 1.2±0.3 0.3±0.1 1.2±0.3 0.6±0.2 0.3±0.1 1.2±0.4 13.0±2.9 | 12.45 12.59 21.40 12.16 11.47 24.41 15.75 140.00 17.25 63.50 149.33 35.92 0.40 |
| 7m 7n 7o 7p 7q 7r 8 9 | D-Phe-OMe Thr-OMe L-Phe-OMe Cys-OMe 2-Cl-Ala-OMe Asp-(OMe) 2 -OMe -NHOH | 46.2±6.4 51.9±3.9 11.0±3.0 50.3±4.3 50.0±4.2 21.7±2.1 63.4±3.2 3.1±0.7 2.4±0.5 | 1.4±0.5 1.8±0.6 1.3±0.3 1.7±0.5 15.6±2.3 0.5±0.1 10.6±2.1 2.2±0.6 3.4±0.6 | 33.00 28.83 8.46 29.59 3.21 43.40 5.98 1.41 0.71 |
| Ubenimex | ||||
Numerical value is the mean value of three experiments in a table, " ± " back numeric representation standard deviation.
Embodiment 8 target compounds suppress lotus liver cancer H22 mouse blood road and shift test (in vivo test)
Concrete grammar and operation steps are seen CN 1528745A " pyrrolidinyl metalloprotease inhibitor and preparation method thereof "
Table two, in vivo test result
| Compound | Mouse survival number (n) | Body weight (g) | Lung heavy (g) | Lung surface tubercle number (n) | Inhibiting rate (%) |
| Blank 7a 7g 7h 7i 7j 7k 7l 7m 7n 7r 89 | 10 10 10 9(10) * 10 10 10 9(10) * 10 10 9(10) * 10 10 | 22.70±3.45 21.03.±4.64 19.80±3.72 20.65±2.32 22.20±2.66 23.65±3.31 20.54±2.42 21.12±2.67 20.22±4.26 23.10±4.04 21.00±1.62 23.48±3.50 20.17±3.35 | 0.170±0.020 O.172±0.039 0.153±0.031 0.163±0.018 0.173±0.014 0.154±0.022 O.168±0.020 0.175±0.042 0.169±O.019 0.172±O.042 0.164±0.089 0.163±0.033 0.165±0.032 | 47.30±2.01 15.67±3.78 9.35±4.24 17.5±0.86 13.2±0.78 8.34±2.34 10.6±1.32 9.03±2.63 15.16±1.72 11.46±0.57 11.23±2.43 24.3±1.68 7.25±1.52 | - 66.87 80.23 63.00 72.09 82.37 77.59 80.91 67.95 75.77 76.26 48.63 84.67 |
*Be the initial number of animals of experiment in ()
When data processing adopted One-Way ANOVA method comparative group differences, each organized body weight and blank group there are no significant difference (p＜0.05); Each organizes the heavy and blank group of lung there are no significant difference (p＜0.05); There is utmost point significant difference (p＜0.01) in the tubercle number average.Compare with the blank group, the heavy there was no significant difference explanation of body weight and lung institute synthetic class inhibitor peptides toxic side effect is less; The tubercle number obviously reduces explanation institute synthetic class inhibitor peptides and has shown inhibition metastases activity in the fabulous body.
Claims (10)
1. the class peptide compounds that has general formula I, with and optical isomer, diastereomer and racemic mixture, its pharmacy acceptable salt, solvate or prodrug,
Wherein,
R
1Be aryl, heteroaryl, aryl C1-6 alkyl, heteroaryl C1-9 alkyl, aryl C2-6 thiazolinyl, heteroaryl C2-6 thiazolinyl, aryl C2-6 alkynyl, heteroaryl C2-6 alkynyl, the C1-6 alkyl, the N end is protected or is not protected natural or the alpha-non-natural amino acid derivative, optional by one or more following groups replacements: hydroxyl, halogen, nitro, cyano group, halogen C1-8 alkyl, C1-8 alkoxyl group, the C1-6 alkyl-carbonyl, C1-8 carbalkoxy, aryl C1-8 carbalkoxy;
R
2Be hydrogen, aryl, heteroaryl, aryl C1-6 alkyl, heteroaryl C1-9 alkyl, aryl C2-6 thiazolinyl, heteroaryl C2-6 thiazolinyl, aryl C2-6 alkynyl, heteroaryl C2-6 alkynyl, and various natural or alpha-non-natural amino acid side chains, optional by one or more following groups replacements: halogen, nitro, cyano group, halogen C1-8 alkyl, C1-8 alkoxyl group, the C1-6 alkyl-carbonyl, the C1-8 carbalkoxy;
R
3Be OH, NHOR
10(R here
10Be hydrogen or C1-8 alkyl) the C1-8 alkoxyl group, NHR
11(R here
11Be the C1-8 alkyl), aryl, heteroaryl, aryl C1-6 alkyl, heteroaryl C1-9 alkyl, aryl C2-6 thiazolinyl, heteroaryl C2-6 thiazolinyl, aryl C2-6 alkynyl, heteroaryl C2-6 alkynyl, NHCH (R
2) COR
4(R here
4Be OH or NHOR
10), optional by one or more following groups replacements: halogen, nitro, cyano group, halogen C1-8 alkyl, the C1-8 alkoxyl group, the C1-6 alkyl-carbonyl, C1-8 alcoxyl carbonyl,
N is 1 or 2,
*Be that steric configuration is S or R optical purity or its raceme.
2. class peptide compounds as claimed in claim 1 is characterized in that:
R
1Be aryl, aryl C1-6 alkyl, aryl C2-6 thiazolinyl, the non-steroidal anti-inflammatory drugs organic acid, the N end is protected or do not protected natural or the alpha-non-natural amino acid derivative, and is optional by one or more following groups replacements: hydroxyl, halogen, nitro, cyano group, C1-8 alkoxyl group;
R
2Be hydrogen, and various natural or alpha-non-natural amino acid side chain;
R
3Be OH, OCH
3, NHCH (R
2) COR
4, R here
4Be OCH
3Or NHOH.
3. as each compound of claim 1-2, it is characterized in that it being following compound:
(S)-methyl-2-(2-(2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) acetic ester,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3 Methylbutanoic acid ester,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-4-methylpent acid esters,
(2S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3 methylvaleric acid ester,
(S)-methyl 1-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) ethanoyl) tetramethyleneimine-2-carboxylicesters,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) propionic ester,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-5-(3-nitro guanidine radicals) valerate,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-4-(methylthio group) butyric ester,
(S)-methyl 6-(benzyloxy carbonyl amide group)-2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) capronate,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-(4-hydroxybenzyl) propionic ester,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-(1H-indol-3-yl) propionic ester,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-(1H-imidazoles-5-yl) propionic ester,
(R)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-phenylpropionic acid ester,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-hydroxy propionate,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-phenylpropionic acid ester,
(S)-methyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido)-3-mercaptopropionic acid ester,
(S)-dimethyl 2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) phthalic ester,
(S)-methyl 3-chloro-2-(2-((S)-2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) propionic ester,
(S)-methyl 3-(2-(2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetamido) propionic ester,
(S)-methyl 2-(2,6-dioxy-3-(3,4,5-trimethoxy-benzene amido) piperidines-1-yl) acetic ester,
(S)-and N-(1-(2-(azanol base)-2-oxygen ethyl)-2,6-dioxopiperidin-3-yl)-3,4,5-trimethoxy aniline.
4. the intermediate of the described class peptide compounds of preparation claim 1 is characterized in that, is 3,4, the 5-trimethoxybenzoic acid; 3,4, the 5-trimethoxy-benzoyl chloride; (S)-2-(3,4,5-trimethoxy anilino) pentanedioic acid; (S)-and N-(2,6-dioxy-tetrahydrochysene-2H-pyrans-3-yl)-3,4, the 5-trimethoxy-benzamide; (S)-2-(2,6-dioxy-3-(3,4,5-trimethoxy benzamido) piperidines-1-yl) acetate, (S)-tertiary butyl-2,6-dioxopiperidin-3-aminocarbamic acid ester, (S)-ethyl-2-(3-(t-butoxycarbonyl amino)-2,6-dioxopiperidin-1-yl) acetic ester, (S)-benzyl-2,6-dioxopiperidin-3-aminocarbamic acid ester, (S)-ethyl 2-(3-(benzyloxycarbonyl amino)-2,6-dioxopiperidin-1-yl) acetic ester.
5. the preparation method of the described class peptide compounds of claim 1 is characterized in that, is raw material with the optical purity acidic amino acid, condensation under alkaline condition, the compound of gained passes through dehydration condensation again, nucleophilic substitution, and polypeptide condensation reagent condensations etc. are reacted final separation and purification and are got; Or be raw material with optically pure glutamine or l-asparagine; by the synthetic key intermediate of steps such as Boc or Cbz protection, cyclization, replacement; deprotection then, class dipeptides or class tripeptides again by obtaining different series with the different acyl chlorides and the amino acid derivative of biologically active.
6. each class peptide compounds of claim 1-3 comprises matrix metalloproteinase or Aminopeptidase N at prevention or treatment and metalloprotease, the application of the medicine of the mammalian diseases that active unconventionality expression is relevant.
7. application as claimed in claim 6, the described and related mammalian disease metal proteinase activity unconventionality expression comprises: inflammation, cancer, multiple sclerosis, various tissue ulcers or tissue ulcer's venereal disease disease, periodontopathy, epidermolysis bullosa, leukemia etc.
8. pharmaceutical composition comprises each class peptide compounds and (2) one or more pharmaceutically acceptable carriers or vehicle of (1) claim 1-3.
9. one kind is suitable for the mammiferous pharmaceutical composition of orally give, comprises the arbitrary class peptide compounds of (1) claim 1-3 and (2) pharmaceutically acceptable carrier, optional (3) one or more the pharmaceutically acceptable vehicle that comprise.
10. one kind is suitable for parenteral and gives mammiferous pharmaceutical composition, comprises the arbitrary class peptide compounds of (1) claim 1-3 and (2) pharmaceutically acceptable carrier, optional (3) one or more the pharmaceutically acceptable vehicle that comprise.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101357893B (en) * | 2008-08-22 | 2011-12-07 | 山东大学 | Ethylene diamine metalloid protease inhibitor and use thereof |
| CN101538311B (en) * | 2009-04-16 | 2012-10-31 | 山东大学 | Alpha-amido acyl-ring imide peptoid metalloprotease inhibitor and application thereof |
| CN103848778A (en) * | 2014-03-28 | 2014-06-11 | 潍坊高新生物园发展有限公司 | Aminopeptidase N inhibitor and preparation method and application thereof |
| CN113801009A (en) * | 2021-09-26 | 2021-12-17 | 武汉工程大学 | Method for catalyzing gallic acid methylation by using ionic liquid |
-
2006
- 2006-12-12 CN CNB2006101462075A patent/CN100560568C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101357893B (en) * | 2008-08-22 | 2011-12-07 | 山东大学 | Ethylene diamine metalloid protease inhibitor and use thereof |
| CN101538311B (en) * | 2009-04-16 | 2012-10-31 | 山东大学 | Alpha-amido acyl-ring imide peptoid metalloprotease inhibitor and application thereof |
| CN103848778A (en) * | 2014-03-28 | 2014-06-11 | 潍坊高新生物园发展有限公司 | Aminopeptidase N inhibitor and preparation method and application thereof |
| CN103848778B (en) * | 2014-03-28 | 2016-01-06 | 潍坊高新生物园发展有限公司 | A kind of aminopeptidase N inhibitor and preparation method thereof and application |
| CN113801009A (en) * | 2021-09-26 | 2021-12-17 | 武汉工程大学 | Method for catalyzing gallic acid methylation by using ionic liquid |
| CN113801009B (en) * | 2021-09-26 | 2022-12-20 | 武汉工程大学 | Method for catalyzing gallic acid methylation by using ionic liquid |
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